Recombinant Anti-GNAQ antibody [EPR20978] (ab210004)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20978] to GNAQ
- Suitable for: IP, IHC-P, WB, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-GNAQ antibody [EPR20978]
See all GNAQ primary antibodies -
Description
Rabbit monoclonal [EPR20978] to GNAQ -
Host species
Rabbit -
Specificity
The sequence homology between GNAQ and three other related Ga family members: GNA11, GNA14 and GNA15 is 90.3%, 79.9% and 53.5%, respectively. -
Tested applications
Suitable for: IP, IHC-P, WB, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293T, HeLa, A549, Jurkat, RAW 264.7, PC-12 and C6 whole cell lysates; Human stomach and fetal pancreas lysates; Mouse lung and kidney lysates; Rat lung lysate; His-tagged human GNAQ (aa1-359) recombinant protein. IHC-P: Human mammary gland and papillary thyroid carcinoma tissues. Flow Cyt (intra): HEK-293T cells. IP: A549 whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20978 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab210004 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IP |
1/30.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This antibody is not recommended for mouse and rat species in IHC application. Under our experiment conditions this antibody showed weak or no staining on mouse and rat tissues. |
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WB |
1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
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Flow Cyt (Intra) |
1/50.
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Notes |
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IP
1/30. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. This antibody is not recommended for mouse and rat species in IHC application. Under our experiment conditions this antibody showed weak or no staining on mouse and rat tissues. |
WB
1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa). |
Flow Cyt (Intra)
1/50. |
Target
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Function
Guanine nucleotide-binding proteins (G proteins) are involved as modulators or transducers in various transmembrane signaling systems. -
Tissue specificity
Predominantly expressed in ovary, prostate, testis and colon. -
Sequence similarities
Belongs to the G-alpha family. G(q) subfamily. - Information by UniProt
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Database links
- Entrez Gene: 2776 Human
- Entrez Gene: 14682 Mouse
- Entrez Gene: 81666 Rat
- Omim: 600998 Human
- SwissProt: P50148 Human
- SwissProt: P21279 Mouse
- SwissProt: P82471 Rat
- Unigene: 269782 Human
see all -
Alternative names
- CMC1 antibody
- G alpha Q antibody
- G protein alpha Q antibody
see all
Images
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: GNAQ knockout HAP1 whole cell lysate (20 µg)Lanes 1 - 2: Merged signal (red and green). Green - ab210004 observed at 42 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab210004 was shown to specifically react with GNAQ in wild-type HAP1 cells as signal was lost in GNAQ knockout cells. Wild-type and GNAQ knockout samples were subjected to SDS-PAGE. ab210004 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution
Lane 1 : His-tagged human GNAQ (aa1-359) recombinant protein
Lane 2 : His-tagged human GNA11 (aa1-359) recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 1 secondBlocking and dilution buffer: 5% NFDM/TBST
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Lanes 1-4 : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/2000 dilution
Lanes 5-7 : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution
Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 3 : A549 (human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 5 : C6 (rat glial tumor cell line) whole cell lysate at 10 µg
Lane 6 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 7 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 42 kDa
Observed band size: 42 kDaExposure time : Lanes 1-3 & 5-7: 3 minutes; Lane 4: 3 seconds.
Blocking and dilution buffer: 5% NFDM/TBST
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Lanes 1-4 : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/2000 dilution
Lane 5 : Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution
Lane 1 : Human fetal pancreas lysate at 20 µg
Lane 2 : Mouse lung lysate at 20 µg
Lane 3 : Mouse kidney lysate at 20 µg
Lane 4 : Rat lung lysate at 20 µg
Lane 5 : Human stomach lysate at 10 µg
Secondary
Lanes 1-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 5 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Developed using the ECL technique.
Predicted band size: 42 kDa
Observed band size: 42 kDaExposure time : Lanes 2-4: 3 minutes; Lanes 1 & 5: 15 seconds.
Blocking and dilution buffer: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAQ antibody [EPR20978] (ab210004)
Immunohistochemical analysis of paraffin-embedded human mammary gland tissue labeling GNAQ with ab210004 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on human mammary gland is observed (PMID: 22421440; PMID: 1946421; PMID: 18799799). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAQ antibody [EPR20978] (ab210004)
Immunohistochemical analysis of paraffin-embedded human papillary thyroid carcinoma tissue labeling GNAQ with ab210004 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on human mammary gland is observed (PMID: 22421440; PMID: 1946421; PMID: 18799799). Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methonol-permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell line labeling GNAQ with ab210004 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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GNAQ was immunoprecipitated from 0.35 mg of A549 (human lung carcinoma cell line) whole cell lysate with ab210004 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab210004 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: A549 whole cell lysate 10 μg (Input).
Lane 2: ab210004 IP in A549 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab210004 in A549 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (2)
ab210004 has been referenced in 2 publications.
- Govatati S et al. Myristoylation of LMCD1 Leads to Its Species-Specific Derepression of E2F1 and NFATc1 in the Modulation of CDC6 and IL-33 Expression During Development of Vascular Lesions. Arterioscler Thromb Vasc Biol 40:1256-1274 (2020). PubMed: 32160773
- Govatati S et al. NFATc1-E2F1-LMCD1-Mediated IL-33 Expression by Thrombin Is Required for Injury-Induced Neointima Formation. Arterioscler Thromb Vasc Biol 39:1212-1226 (2019). PubMed: 31043075